Department of Surgical Oncology and Technology, Imperial College London, St Mary's Hospital, London, United Kingdom.
J Surg Res. 2010 Dec;164(2):e265-72. doi: 10.1016/j.jss.2010.09.041. Epub 2010 Oct 16.
Peritoneal metastasis occurs in up to 30% of patients with gastric cancer. The aim of this experimental study is to develop and validate a novel ex vivo model of the human peritoneum to better identify factors involved in the development of peritoneal metastasis in order to improve its management and prognosis.
Peritoneal discs harvested from hernia sacs obtained at inguinal hernia surgery were suspended in media using Teflon rings. Viability of the tissue was investigated using MTS assay, light and scanning electron microscopy (LM and SEM) over 72 h. To assess validity of the model, phenotypic changes in tumor cells were investigated. Changes in matrix metalloproteinases (MMP)-2 and -9 activities in HGC and AGS gastric adenocarcinoma cells after co-culture were investigated using zymography. Modulation of tumor cell adhesion to peritoneum after exposure to heparin was assessed using a fluorometric adhesion assay. Analysis was performed using Kruskal-Wallis for multiple comparisons and Mann-Witney U for comparisons between each group.
MTS assay showed reduced viability after 72 h (P = 0.047, compared with 24 h). Mesothelial cell loss at 48 h was demonstrated by LM and SEM, confirming peritoneal viability for at least 24 h after tissue harvesting. Zymography confirmed increased MMP2 and -9 activities in tumor cells and peritoneal tissue during co-culture compared with controls, and heparin significantly reduced tumor cell adherence (P = 0.04), as observed in published in vivo models.
A validated complete model of peritoneum was developed that has shown potential to determine realistic mechanisms of peritoneal metastasis.
腹膜转移发生在高达 30%的胃癌患者中。本实验研究的目的是开发和验证一种新的人腹膜体外模型，以更好地识别参与腹膜转移发展的因素，从而改善其管理和预后。
使用特氟隆环将从腹股沟疝手术获得的疝囊中采集的腹膜片悬挂在培养基中。使用 MTS 测定法、光镜和扫描电子显微镜（LM 和 SEM）在 72 小时内研究组织的活力。为了评估模型的有效性，研究了肿瘤细胞的表型变化。使用明胶酶谱法研究共培养后 HGC 和 AGS 胃腺癌细胞中基质金属蛋白酶（MMP）-2 和 -9 活性的变化。使用荧光吸附测定法评估肝素暴露后肿瘤细胞对腹膜的粘附能力的调节。使用 Kruskal-Wallis 进行多组比较，使用 Mann-Whitney U 进行每组之间的比较。
MTS 测定法显示 72 小时后活力降低（P = 0.047，与 24 小时相比）。LM 和 SEM 显示，48 小时时间皮细胞丢失，证实组织采集后至少 24 小时腹膜仍具有活力。明胶酶谱法证实共培养中肿瘤细胞和腹膜组织的 MMP2 和 -9 活性增加，肝素显著降低肿瘤细胞粘附（P = 0.04），如已发表的体内模型所示。
开发了一种经过验证的完整腹膜模型，该模型显示出确定腹膜转移真实机制的潜力。
